Real-Time Monitoring of a Single Molecule in Sub-nanometer Space by Dynamic Surface-Enhanced Raman Spectroscopy

In biology and chemistry, the ultimate goal is to monitor single molecules without labels. However, long-term monitoring of label-free molecules remains a challenge. Here, on the basis of the photothermal effect of gold nanorods (GNRs), we developed a platform for monitoring of a single molecule employing surface-enhanced Raman spectroscopy (SERS). Laser re-irradiation forms 1.0 nm gaps between GNRs, allowing us to observe single crystal violet (CV) molecules blinking for up to 4 min with dynamic surface-enhanced Raman spectroscopy (D-SERS). Bianalyte experiments confirm single-molecule features at CV concentrations of 10^-14 M. Combining density functional theory (DFT) calculations with a free CV molecule observed in millisecond D-SERS, we propose that CV molecules can be confined to sub-nanometer space and the orientation of an individual CV moving in the range of 50-90° can be dynamically captured by D-SERS. This will provide a novel idea for effective exploration of the temporal and spatial dynamic processes of different reactions.